Continuous cultivation method of Aspergillus niger citrate seeds based on mycelium ball dispersion technology

A technology of mycelium balls and citric acid, applied in the field of fermentation engineering, can solve the problems of high raw material cost and operating cost, complex preparation process of Aspergillus niger spores, and heavy screening workload, so as to reduce raw material consumption cost and operating cost, reduce Mature seed culture time, reducing the effect of raw material and energy consumption

Active Publication Date: 2016-07-06
JIANGNAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional seed culture process uses the inoculation method of Aspergillus niger spores, which requires a large number of mature Aspergillus niger spores, and the preparation process of Aspergillus niger spores is relatively complicated
A batch of mature Aspergillus niger spores needs to be screened on plates, cultivated on inclined planes, cultivated in eggplant bottles, and finally cultured in bran koji tanks. The preparation process is cumbersome, the activity screening is complicated, the workload is large, and the preparation cycle needs to be more than 30 days.
At the same time, the process of cultivating seeds by spore inoculation only requires at least 12 hours for spore germination, so mature seeds take a long time to cultivate
[0004] The traditional step-by-step expansion of Aspergillus niger spores has disadvantages such as long and complicated process, heavy screening workload, long cycle, etc., which consumes a lot of raw material costs and operating costs

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Glucose and soybean meal powder were used to prepare seed medium (8% total sugar, 0.15% total nitrogen) and fermentation medium (16% total sugar, 0.15% total nitrogen) respectively; the concentration of spores after transplanting was 550,000 / ml Inoculation: Cultivate for 20 hours to obtain mature seed liquid, the mycelium of the seed liquid is dispersed by a disperser, and the average diameter of the treated bacterial balls is 100um, which are respectively inserted into the next-level seed medium and fermentation medium, and the inoculation amount is 5% . Among them, those who are connected to the next-level seed medium, the seeds are cultivated for 24 hours, the seed liquid is dispersed by a disperser, and the average diameter of the treated bacterial balls is 80um, which are respectively inserted into the next-level seed medium and fermentation medium; For the first-grade fermentation medium, when the reducing sugar concentration in the fermentation medium drops below...

Embodiment 2

[0027] Mix corn flour and deionized water in a proportion of 1:3 in a batching tank at about 60°C, add calcium hydroxide to adjust the pH to 6.0, add high-temperature α-amylase at 25U / g corn flour, and liquefy through secondary spraying , passed the iodine test (light brown) to obtain a corn liquefaction mixture; 80% of the mixture was filtered through a plate frame to obtain a corn liquefaction clear liquid. The corn liquefaction mixture and clear liquid were mixed in a certain proportion to prepare seed medium (10% total sugar, add a certain amount of ammonium sulfate to adjust total nitrogen 0.4%) and fermentation medium (14% total sugar, 0.05% total nitrogen). After transplanting, the concentration of the spore liquid is 580,000 / ml for inoculation; after culturing for 26 hours, mature seed liquid is obtained, and the mycelium of the seed liquid is dispersed by a disperser. Base and fermentation medium, the inoculum size was 15% (v / v) and 10% (v / v) respectively. Wherein th...

Embodiment 3

[0030] Same as embodiment 2, corn flour and water are mixed evenly according to a certain ratio, and the corn liquefaction mixed solution is obtained through secondary jet liquefaction, the corn liquefaction mixed solution is mixed with water according to a certain ratio, and a certain amount of ammonium sulfate is added to prepare the seed culture medium (total sugar 8%, total nitrogen 0.2%). Mix cassava flour and water in a proportion of 1:4 in a batching tank at about 60°C, add calcium hydroxide to adjust the pH to 5.8, add high-temperature α-amylase at 25U / g cassava flour, and liquefy through secondary spraying. The iodine test is qualified, and the cassava liquefaction liquid is obtained; the cassava liquefaction liquid is mixed with water according to a certain ratio, and a certain amount of ammonium sulfate is added to prepare a fermentation medium (total sugar 15.6%, total nitrogen 0.076%). After transplanting, the concentration of the spore liquid was 480,000 / ml for i...

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Abstract

A citric acid aspergillus niger seed continuous culture method based on a mycelium pellet dispersion technology comprises the steps as follows: (1), an aspergillus niger spore solution is inoculated to a seed culture medium and cultured and prepared into a mature seed solution; (2), the mature seed solution is treated by a disperser to obtain a dispersed mycelium seed solution; (3), part of the dispersed mycelium seed solution obtained in the step (2) is transferred to a fermentation medium for fermentation with 5%-15% of inoculum concentration, and the fermentation is finished when the reducing sugar concentration of the fermentation medium is lower than 0.5%; and (4), part of the dispersed mycelium seed solution obtained in the step (2) is inoculated to a seed culture medium with 5%-15% of inoculum concentration to be cultured and prepared into a next level of mature seed solution, and then the operation returns to the step (2), so that the continuous culture for aspergillus niger seeds is realized. According to the aspergillus niger seed continuous culture method provided by the invention, the tedious culture process of aspergillus niger spores can be directly reduced, simultaneously, the culture time of mature seeds can be effectively shortened, and the utilization rate of raw materials is increased.

Description

technical field [0001] The invention relates to the technical field of fermentation engineering, in particular to a continuous cultivation method for Aspergillus niger citrate seeds based on mycelium ball dispersion technology. Background technique [0002] Citric acid (CitricAcid) also known as citric acid, chemical name is 2-hydroxypropanetricarboxylic acid, colorless crystal, often contains a molecule of crystal water, odorless, has a strong sour taste, soluble in water, in cold water It is easier to dissolve than hot water and is often used for identification and separation. It has a wide range of uses in food, cosmetics, medicine, construction, printing and dyeing, washing and other industries. Because of its pleasant sour taste, refreshing taste, no aftertaste, safety and non-toxicity, it has become the most produced and consumed food in the world. organic acids. The demand for citric acid in emerging industries is showing a gradual increase. Citric acid has a wide ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12P7/48C12R1/685
Inventor 石贵阳陈坚王宝石张杰胡志杰蒋小东孙福新李赢张梁李由然丁重阳顾正华
Owner JIANGNAN UNIV
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